Quantitative assessment of leading edge adhesion: reattachment kinetics modulated by injury-derived intracellular calcium predict wound closure rates in endothelial monolayers

Journal of Cellular Physiology
G M UngerP J Sammak

Abstract

Migrating cells continually develop new substrate attachments at the leading edge (LE) in order to maintain traction for movement. This study evaluates the relationship between LE adhesion and wound closure by modulating injury-derived intracellular free Ca2+ ([Ca2+]i) signaling in endothelial cell (EC) monolayers following scrape-wounding. These data show that brief treatment with increased extracellular Ca2+ ([Ca2+]e) during wounding accelerated wound area closure rates by 50-65%, while brief treatments with calcium influx inhibitors reduced rates by 30-50%. Fura-2 studies in wounded monolayers indicated supranormal [Ca2+]e during wounding increased (by 52%), while influx-inhibitors decreased (by 36%) the percentage of cells exhibiting elevated plateau [Ca2+]i levels. Quantitative time-lapse interference reflection microscopy (IRM) together with indirect alphavbeta3 integrin immunofluorescence was used to measure the effects of 100 microM Gd3+ and 5 mM [Ca2+]e treatment on fractional LE adhesion after wounding. Influx inhibition blocked development of increased injury-derived LE adhesion. Measurements indicated a linear relationship (r2 = 0.99, 0.98) between LE adhesion, development rates (quantified as an association rate co...Continue Reading

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Citations

Sep 5, 2002·Cell Motility and the Cytoskeleton·Lance A DavidsonRay Keller
Feb 18, 2003·Arteriosclerosis, Thrombosis, and Vascular Biology·Pinaki ChaudhuriLinda M Graham
Nov 18, 2005·Histochemistry and Cell Biology·Keiko FujitaMasumi Akita
Jul 17, 2001·American Journal of Physiology. Heart and Circulatory Physiology·C KimuraY Ito

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